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[Protein expression, purification and mouse antiserum preparation of monkeypox virus A23R].
Wang, Yihao; Li, Mingzhi; Jia, Mengle; Yang, Lingdi; Xiong, Jiaqi; Wang, Ting; Wang, Yu; Liu, Shurong; Guo, Wenli; Kong, Lingbao; Li, Meifeng.
Afiliação
  • Wang Y; Institute of Pathogenic Microorganism, Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Department of biotechnology, College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330045, China.
  • Li M; Institute of Pathogenic Microorganism, Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Department of biotechnology, College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330045, China.
  • Jia M; Institute of Pathogenic Microorganism, Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Department of biotechnology, College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330045, China.
  • Yang L; Institute of Pathogenic Microorganism, Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Department of biotechnology, College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330045, China.
  • Xiong J; Institute of Pathogenic Microorganism, Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Department of biotechnology, College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330045, China.
  • Wang T; Institute of Pathogenic Microorganism, Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Department of biotechnology, College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330045, China.
  • Wang Y; Institute of Pathogenic Microorganism, Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Department of biotechnology, College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330045, China.
  • Liu S; Institute of Pathogenic Microorganism, Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Department of biotechnology, College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330045, China.
  • Guo W; Institute of Pathogenic Microorganism, Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Department of biotechnology, College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330045, China.
  • Kong L; Institute of Pathogenic Microorganism, Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Department of biotechnology, College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330045, China. *Corresponding authors, E-mail: lingbaok@mail.jxau.edu.cn.
  • Li M; Institute of Pathogenic Microorganism, Nanchang City Key Laboratory of Animal Virus and Genetic Engineering, Department of biotechnology, College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang 330045, China. *Corresponding authors, E-mail: meifengli77@jxau.edu.cn.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 39(7): 642-648, 2023 Jul.
Article em Zh | MEDLINE | ID: mdl-37403724
ABSTRACT
Objective To express the monkeypox virus (MPXV) A23R protein in Escherichia coli and purify by Ni-NTA affinity column, and to prepare mouse antiserum against MPXV A23R. Methods The recombinant plasmid pET-28a-MPXV-A23R was constructed and transformed into Escherichia coli BL21 to induce the expression of A23R protein. After optimizing the conditions of expression, A23R protein was highly expressed. Recombinant A23R protein was purified by Ni-NTA affinity column and identified by Western blot analysis. The purified protein was used to immunize mice for preparing the A23R polyclonal antibody, and the antibody titer was detected by ELISA. Results The expression of A23R recombinant protein reached the peak under the induced conditions of 0.6 mmol/L isopropyl-ß-D-thiogalactoside (IPTG), 37 DegreesCelsius and 20 hours. The purity of the protein was about 96.07% and was identified by Western blot analysis. The mice were immunized with recombinant protein, and the titer of antibody reached 1102 400 at the 6th week after immunization. Conclusion MPXV A23R is expressed highly and purified with a high purity and its antiserum from mouse is obtained with a high titre.
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Monkeypox virus / Anticorpos Limite: Animals Idioma: Zh Revista: Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi Assunto da revista: ALERGIA E IMUNOLOGIA Ano de publicação: 2023 Tipo de documento: Article País de afiliação: China
Buscar no Google
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Monkeypox virus / Anticorpos Limite: Animals Idioma: Zh Revista: Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi Assunto da revista: ALERGIA E IMUNOLOGIA Ano de publicação: 2023 Tipo de documento: Article País de afiliação: China